ENGLISH

Cytoskeleton Dynamics

By Helder Maiato (ed.)

Book information

Publisher
Springer
Year
2020
ISBN
978-1-0716-0218-8, 978-1-0716-0219-5
Language
english
Format
PDF
Filesize
12 MB (12949863 bytes)
Series
Methods and Protocols 2101
Pages
390\389
Topic
Biology Molecular
Time added
2019-12-27 18:19:47

Description

This volume details comprehensive state-of-the-art methods on actin microfilaments and microtubules and how they work to achieve different cellular functions in different cellular contexts. Chapters guide readers through protein purification, in vitro reconstitution of several cytoskeleton properties, analyses of microtubule- and actin-based structures, functional dissection of post-translational modifications, and roles in several biological processes. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Cytoskeleton Dynamics: Methods and Protocols aims to provide a wide range of experimental approaches and be an invaluable resource for present and future generations of cytoskeleton researchers. Front Matter ....Pages i-xi Purification of Mammalian Tubulins and Tubulin-Associated Proteins Using a P2A-Based Expression System (Nuo Yu, Niels Galjart)....Pages 1-17 Expression and Purification of Microtubule-Associated Proteins from HEK293T Cells for In Vitro Reconstitution (Shasha Hua, Kai Jiang)....Pages 19-26 In Vitro Reconstitution Assays of Microtubule Amplification and Lattice Repair by the Microtubule-Severing Enzymes Katanin and Spastin (Annapurna Vemu, Ewa Szczesna, Antonina Roll-Mecak)....Pages 27-38 In Vitro Microtubule Dynamics Assays Using Dark-Field Microscopy (Jeffrey O. Spector, Annapurna Vemu, Antonina Roll-Mecak)....Pages 39-51 In Vitro Reconstitution of Dynamic Co-organization of Microtubules and Actin Filaments in Emulsion Droplets (Kim J. A. Vendel, Celine Alkemade, Nemo Andrea, Gijsje H. Koenderink, Marileen Dogterom)....Pages 53-75 Studying Tau-Microtubule Interaction Using Single-Molecule TIRF Microscopy (Virginie Stoppin-Mellet, Nassiba Bagdadi, Yasmina Saoudi, Isabelle Arnal)....Pages 77-91 In Vitro FRET- and Fluorescence-Based Assays to Study Protein Conformation and Protein-Protein Interactions in Mitosis (Stephanie C. Ems-McClung, Claire E. Walczak)....Pages 93-122 Methods for Studying Membrane-Associated Bacterial Cytoskeleton Proteins In Vivo by TIRF Microscopy (Charlène Cornilleau, Arnaud Chastanet, Cyrille Billaudeau, Rut Carballido-López)....Pages 123-133 Processing TIRF Microscopy Images to Characterize the Dynamics and Morphology of Bacterial Actin-Like Assemblies (Cyrille Billaudeau, Arnaud Chastanet, Rut Carballido-López)....Pages 135-145 Studying Centriole Duplication and Elongation in Human Cells (Catarina Peneda, Carla A. M. Lopes, Mónica Bettencourt-Dias)....Pages 147-162 Assaying Microtubule Nucleation (Artur Ezquerra, Ricardo Viais, Jens Lüders)....Pages 163-178 Detection of Microtubule Nucleation Hotspots at the Golgi (Roslin J. Thoppil, Anna A. W. M. Sanders, Irina Kaverina)....Pages 179-189 Ultrastructural Analysis of Microtubule Ends (J. Richard McIntosh, Eileen O’Toole, Cynthia Page, Garry Morgan)....Pages 191-209 Optogenetic Control of Microtubule Dynamics (Jeffrey van Haren, Lauren S. Adachi, Torsten Wittmann)....Pages 211-234 Measurement of Microtubule Half-Life and Poleward Flux in the Mitotic Spindle by Photoactivation of Fluorescent Tubulin (Hugo Girão, Helder Maiato)....Pages 235-246 Functional Dissection of Mitosis Using Immortalized Fibroblasts from the Indian Muntjac, a Placental Mammal with Only Three Chromosomes (Ana C. Almeida, Danica Drpic, Naoyuki Okada, Joana Bravo, Marta Madureira, Helder Maiato)....Pages 247-266 Visualization and Functional Analysis of Spindle Actin and Chromosome Segregation in Mammalian Oocytes (Binyam Mogessie)....Pages 267-295 Using the Four-Cell C. elegans Embryo to Study Contractile Ring Dynamics During Cytokinesis (Fung Yi Chan, Ana Marta Silva, Ana Xavier Carvalho)....Pages 297-325 Knocking Out Multiple Genes in Cultured Primary Neurons to Study Tubulin Posttranslational Modifications (Satish Bodakuntla, Carsten Janke, Maria M. Magiera)....Pages 327-351 Measuring the Impact of Tubulin Posttranslational Modifications on Axonal Transport (Satish Bodakuntla, Maria M. Magiera, Carsten Janke)....Pages 353-370 Methods to Measure Perinuclear Actin Dynamics During Nuclear Movement in Migrating Cells (Cátia S. Janota, Francisco J. Calero-Cuenca, Edgar R. Gomes)....Pages 371-385 Back Matter ....Pages 387-390

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